First report on prevalence, molecular characterization and phylogenetic study of Toxoplasma gondii infecting sheep of the Malakand Division of Pakistan.

BACKGROUND: Toxoplasma gondii is an apicomplexan protozoan parasite that infects one-third of the population of the world, including humans, animals, birds, and other vertebrates. The present investigation is the first molecular attempt in the Malakand Division of Pakistan to determine the epidemiology and phylogenetic study of Toxoplasma gondii infecting small ruminants. METHODOLOGY: A total of (N = 450) blood samples of sheep were randomly collected during the study period (December 2020 to November 2021), and DNA detection was done using PCR by amplifying ITS-1 genes. SPSS.20 and MEGA-11 software were used for statistical significance and phylogenetic analysis. RESULTS: The overall prevalence of T. gondii infection among sheep was 14.44 % (65/450). A high infection rate was found in more than five-year-olds at 18.33 % (11/60). Sequencing and BLAST analysis of PCR-positive samples confirmed the presence of T. gondii. Randomly, three isolates were sequenced and submitted to GenBank under accession numbers (PP028089-PP028091), respectively. The BLAST analysis of the obtained sequences based on the ITS-1 gene showed 99 % similarities with reported genotypes found in goats of Malakand, Pakistan (PP028089) and dogs of Brazil (MF766454). The study concludes that T. gondii is notably prevalent among the sheep population in the region, emphasizing the significant role of risk factors in disease transmission across animals and potentially to humans. Further research, zoonotic potential analysis, and targeted control measures are warranted to address and manage this parasitic infection effectively.

MicroRNA-129-1-3p protects chicken granulosa cells from cadmium-induced apoptosis by down-regulating the MCU-mediated Ca2+ signaling pathway.

Cadmium (Cd) is known as a female reproductive toxicant. Our previous study has shown that Cd can influence the proliferation and cell cycle of granulosa cells and induce apoptosis. MicroRNAs (miRNAs) play an important role in the regulation of Cd-induced granulosa cell damage in chickens. However, the mechanism remains unclear. In this study, we investigated the mechanisms by which microRNA-129-1-3p (miR-129-1-3p) regulates Cd-induced cytotoxicity in chicken granulosa cells. As anticipated, exposure to Cd resulted in the induction of oxidative stress in granulosa cells, accompanied by the downregulation of antioxidant molecules and/or enzymes of Nrf2, Mn-SOD, Cu-Zn SOD and CAT, and the upregulation of Keap1, GST, GSH-Px, GCLM, MDA, hydrogen peroxide and mitochondrial reactive oxygen species (mtROS). Further studies found that Cd exposure causes mitochondrial calcium ions (Ca2+) overload, provoking mitochondrial damage and apoptosis by upregulating IP3R, GRP75, VDAC1, MCU, CALM1, MFF, caspase 3, and caspase 9 gene and/or protein expressions and mitochondrial Ca2+ levels, while downregulating NCX1, NCLX and MFN2 gene and/or protein expressions and mitochondrial membrane potential (MMP). The Ca2+ chelator BAPTA-AM or the MCU inhibitor MCU-i4 significantly rescued Cd-induced mitochondrial dysfunction, thereby attenuating apoptosis. Additionally, a luciferase reported assay and western blot analysis confirmed that miR-129-1-3p directly target MCU. MiR-129-1-3p overexpression almost completely inhibited protein expression of MCU, increased the gene and protein expressions of NCLX and MFN2 downregulated by Cd, and attenuated mitochondrial Ca2+ overload, MMP depression and mitochondria damage induced by Cd. Moreover, the overexpression of miR-129-1-3p led to a reduction in mtROS and cell apoptosis levels, and a suppression of the gene and protein expressions of caspase 3 and caspase 9. As above, these results provided the evidence that IP3R-MCU signaling pathway activated by Cd plays a significant role in inducing mitochondrial Ca2+ overload, mitochondrial damage, and apoptosis. MiR-129-1-3p exerts a protective effect against Cd-induced granulosa cell apoptosis through the direct inhibition of MCU expression in the ovary of laying hens.

A novel injectable sericin hydrogel with strong fluorescence for tracing.

Hydrogel systems with strong fluorescence, as convenient tracers or bio-probes, have attracted much attention in biomedical engineering. Currently, most hydrogels endowed fluorescent properties due to modifying additional fluorophores. However, these fluorophores owing to photobleaching and toxicity limit the practical applications of hydrogels. Herein, we prepared a novel self-luminescence hydrogel through double crosslinking glutaraldehyde and hydrogen peroxide/horseradish peroxidase (H2O2/HRP) with sericin protein. The double cross-linked sericin hydrogel exhibits strong green and red intrinsic fluorescence which can be excited over a wide range of wavelengths. Moreover, this hydrogel with strong intrinsic fluorescence could penetrate thick pigskin tissue, which has potential application in implantable bio-tracer areas. In addition to the above unique properties, this sericin hydrogel possesses two types of micropore structures with high porosity, swelling properties, pH-responsive degradability, super elasticity, injectability, viscosity, and excellent biocompatibility. The investigation could significantly expand the scope of protein hydrogels in biomedical applications.

MicroRNA-129-1-3p attenuates autophagy-dependent cell death by targeting MCU in granulosa cells of laying hens under H2O2-induced oxidative stress.

The present study aimed to investigate the mechanism of microRNA-129-1-3p (miR-129-1-3p) in regulating hydrogen peroxide (H2O2)-induced autophagic death of chicken granulosa cell by targeting mitochondrial calcium uniporter (MCU). The results indicated that the exposure of hens' ovaries to H2O2 resulted in a significant elevation in reactive oxygen species (ROS) levels, as well as the apoptosis of granulosa cells and follicular atresia. This was accompanied by an upregulation of glucose-regulated protein 75 (GRP75), voltage-dependent anion-selective channel 1 (VDAC1), MCU, mitochondria fission factor (MFF), microtubule-associated protein 1 light chain 3 (LC3) I, and LC3II expression, and a downregulation of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) and mitofusin-2 (MFN2) expression. In hens' granulosa cells, a luciferase reporter assay confirmed that miR-129-1-3p directly regulates MCU. The induction of oxidative stress through H2O2 resulted in the activation of the permeability transition pore, an overload of calcium, depolarization of the mitochondrial membrane potential, dysfunction of mitochondria-associated endoplasmic reticulum membranes (MAMs), and ultimately, autophagic cell death. The overexpression of miR-129-1-3p effectively mitigated these H2O2-induced changes. Furthermore, miR-129-1-3p overexpression in granulosa cells prevented the alterations induced by H2O2 in the expression of key proteins that play crucial roles in maintaining the integrity of MAMs and regulating autophagy, such as GRP75, VDAC1, MFN2, PTEN-induced kinase 1 (Pink1), and parkin RBR E3 ubiquitin-protein ligase (Parkin). Together, these in vitro- and in vivo-based experiments suggest that miR-129-1-3p protects granulosa cells from oxidative stress-induced autophagic cell death by downregulating the MCU-mediated mitochondrial autophagy. miR-129-1-3p/MCU calcium signaling pathway may act as a new target to alleviate follicular atresia caused by oxidative stress in laying hens.

Pseudorogneria libanotica Intraspecific Genetic Polymorphism Revealed by Fluorescence In Situ Hybridization with Newly Identified Tandem Repeats and Wheat Single-Copy Gene Probes.

The genus Pseudoroegneria (Nevski) Löve (Triticeae, Poaceae) with its genome abbreviated 'St' accounts for more than 60% of perennial Triticeae species. The diploid species Psudoroegneria libanotica (2n = 14) contains the most ancient St genome. Therefore, investigating its chromosomes could provide some fundamental information required for subsequent studies of St genome evolution. Here, 24 wheat cDNA probes covering seven chromosome groups were mapped in P. libanotica to distinguish homoelogous chromosomes, and newly identified tandem repeats were performed to differentiate seven chromosome pairs. Using these probes, we investigated intraspecific population chromosomal polymorphism of P. libanotica. We found that (i) a duplicated fragment of the 5St long arm was inserted into the short arm of 2St; (ii) asymmetrical fluorescence in situ hybridization (FISH) hybridization signals among 2St, 5St, and 7St homologous chromosome pairs; and (iii) intraspecific population of polymorphism in P. libanotica. These observations established the integrated molecular karyotype of P. libanotica. Moreover, we suggested heterozygosity due to outcrossing habit and adaptation to the local climate of P. libanotica. Specifically, the generated STlib_96 and STlib_98 repeats showed no cross-hybridization signals with wheat chromosomes, suggesting that they are valuable for identifying alien chromosomes or introgressed fragments of wild relatives in wheat.

Progress on molecular composition and genetic mechanism of plant B chromosomes.

In addition to the standard set of chromosomes (A chromosomes, As), so-called supernumerary B chromosomes (Bs) have been found causing a numerical chromosome variation. Bs have been considered to be genetically inert elements without any functional genes for a long period, because of the limited experimental methods and its dispensable property. More recently, sequencing of dissected Bs from several organisms has revealed the DNA composition, a vast number of protein-coding genes have been found with the effects on the transcripts and protein expression of the host. In this review, we summarize current understanding of B chromosomes carrying plants including rye (Secale cereale L.), maize (Zea mays L.) and Aegilops (Aegilops speltoides Tausch.), with the emphasis on Bs phenotypic effects, the inheritance mechanism of Bs, the molecular composition of Bs, the effects on host transcription regulation and protein expression upon the presence of Bs. Besides, we discuss the current study state and potential application of B chromosomes, aim to provide a new venue for chromosome engineering and breeding research.

MAPK, AKT/FoxO3a and mTOR pathways are involved in cadmium regulating the cell cycle, proliferation and apoptosis of chicken follicular granulosa cells.

The occurrence of cadmium (Cd) in feed is a major problem in animal health and production. Studies have confirmed that Cd depresses egg production of laying hens, which is closely related to follicular atresia. This study aimed to assess the toxic impacts of Cd on the ovarian tissue, and to examine the mechanism of Cd-induced granulosa cell proliferation and apoptosis. Results from the nitric oxide (NO) and malondialdehyde (MDA) content, total superoxide dismutase (T-SOD), glutathione peroxide (GSH-Px), total nitric oxide synthase (T-NOS) and adenosine triphosphatase (ATPase) activities, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, and hematoxylin-eosin (H & E) staining indicated that excess Cd induced oxidative stress, granulosa cell apoptosis and follicular atresia in the layer ovary. Low-dose Cd exposure (1 µM) induced the granulosa cell proliferation, upregulated the mRNA levels of RSK1 and RHEB, activated FoxO3a, AKT, ERK1/2, mTOR and p70S6K1 phosphorylation, and promoted cell cycle progression from phase G1 to S. However, high-dose Cd exposure (15 µM) induced reactive oxygen species (ROS) generation and cell apoptosis, upregulated the mRNA levels of the inflammatory factors, ASK1, JNK, p38 and TAK1, downregulated the expressions of RSK1 and RHEB genes, and inhibited the phosphorylation of ERK1/2, mTOR and p70S6K1 proteins, and the cell cycle progression. Rapamycin pre-treatment completely blocked the phosphorylation of mTOR and p70S6K1 proteins, and the cell cycle progression induced by 1 µM Cd, and accelerated 15 µM Cd-induced cell apoptosis and cell cycle arrest. The microRNA sequencing result showed that 15 µM Cd induced differential expression of microRNA genes, which may regulate AKT, ERK1/2 and mTOR signaling and cell cycle progression by regulating the activity of G proteins and cell cycle-related proteins. Conclusively, these results indicated that Cd can cause the ovarian damage and follicular atresia, and regulate cell cycle, cell proliferation or apoptosis of granulosa cells through MAPK, AKT/FoxO3a and mTOR pathways in laying hens.

Effect of Glycine Nano-Selenium Supplementation on Production Performance, Egg Quality, Serum Biochemistry, Oxidative Status, and the Intestinal Morphology and Absorption of Laying Hens.

The objective of this study was to investigate the feasibility of using glycine nano-selenium (NS-Gly) as a feed supplement and to evaluate its influence on production performance, egg quality, serum biochemistry, oxidative status, and the intestinal morphology and absorption of laying hens. A total of 864 hens at 40 weeks were randomly assigned into six groups including the basal diet (control, 0.13 mg Se/kg); basal diet + 0.30 mg Se/kg (Na2SeO3) diet; and basal diet + 0.15, 0.30, 0.45, and 0.60 mg Se/kg (NS-Gly) diet. After 8 weeks of Se supplementation, no difference was observed among the treatments on production performance and egg quality (P > 0.05). The levels of albumin (ALB) and alanine aminotransferase (GPT) were significantly influenced by dietary Se supplementation (P < 0.05). In the serum, the level of glutathione peroxide (GSH-Px) was significantly increased in the groups with the dietary NS-Gly supplementation (P < 0.05). The superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) levels in all groups of NS-Gly supplementation had a remarkable increase (P < 0.05). In the liver, GSH-Px was significantly increased in 0.45 and 0.60 mg/kg NS-Gly groups (P < 0.05). The activities of SOD and catalase (CAT) were significantly increased in the groups of 0.30 mg/kg NS-Gly diet (P < 0.05). The results of intestinal morphology showed that the crypt depth was affected by higher dose groups of NS-Gly diets in the duodenum, and the differences (P < 0.05) were obtained in villus height, the crypt depth, and the V/C in the jejunum. In the ileum, a significant increase (P < 0.05) of villus height was observed in 0.15 and 0.3 mg/kg Se-added groups. The V/C was the highest in the SS groups (P < 0.05). The mRNA levels of solute carrier family 3 member 1 (rBAT), solute carrier family 6 member 19 (B0AT1), and solute carrier family 15 member 1 (PepT1) increased at different degrees in the duodenum, especially in 0.15 and 0.60 mg/kg NS-Gly groups (P < 0.05). In the jejunum, the expression of B0AT1 was similar to that in the duodenum, and the expression of rBAT increased significantly in the 0.30 and 0.45 mg/kg NS-Gly groups (P < 0.05). The mRNA level of PepT1 increased significantly in the 0.30 mg/kg SS group. Conclusively, dietary NS-Gly supplementation could improve the antioxidant capacity, as well as the structure of small intestine in laying hens, although have no significant effects on the production performance and egg quality.

Dietary High Sodium Fluoride Impairs Digestion and Absorption Ability, Mucosal Immunity, and Alters Cecum Microbial Community of Laying Hens.

(1) Background: This study was conducted to investigate the effects of dietary fluoride (F) on tissue retention, digestive enzymes activities, mucosal immunity, and cecum microbial community of laying hens. (2) Methods: Total of 288 37-week-old Hy-Line Gray laying hens with similar laying rate (85.16% ± 3.87%) were adapted to the basal diets for ten days, and then allocated into three groups at random (n = 9, 6, 6 replicates/group). The concentrations of F in the diets were 31.19 (the control group, CON), 431.38 (F400, low-F group) and 1237.16 mg/kg (F1200, high-F group), respectively. The trial lasted for 59 days. (3) Results: Results suggested that F residuals in duodenum responded to dietary F concentrations positively. The activities of amylase, maltase and lactase were decreased in high-F group, compared with those in the control group. The mRNA expression levels of jejunum and ileum secretory immunoglobulin A (sIgA) and Mucin 2, and sIgA concentrations were decreased inhigh-F group, than those in the control group. The observed operational taxonomic units (OTUs) of laying hens in high-F group were higher than the CON and low-F groups, and the bacterial structure was different from the other two groups. The Lactobacillus was higher in the control group, while Gammaproteobacteria, Escherichia-Shigella, Streptococcaceae, and Enterobacteriaceae were higher in the high-F group. (4) Conclusions: The actual results confirmed that dietary high F intake increased the F residuals in duodenum, and reduced the digestion and absorption of nutrients and immunity via decreasing the activities of digestive enzymes, impairing intestine mucosal immunity, and disturbing the cecum microbial homeostasis of laying hens.

Dietary cadmium chloride impairs shell biomineralization by disrupting the metabolism of the eggshell gland in laying hens.

In this study, we identified cadmium (Cd) as a potential endocrine disruptor that impairs laying performance, egg quality, and eggshell deposition and induces oxidative stress and inflammation in the eggshell glands of laying hens. A total of 480 38-wk-old laying hens were randomly assigned into 5 groups that were fed a basal diet (control) or a basal diet supplemented with Cd (provided as CdCl2·2.5 H2O) at 7.5, 15, 30, and 60 mg Cd per kg feed for 9 wk. The results showed that, when compared with the control group, a low dose of dietary Cd (7.5 mg/kg) had positive effects on egg quality by improving albumen height, Haugh unit, yolk color, and shell thickness at the third or ninth week. However, with the increase in the dose and duration of Cd exposure, the laying performance, egg quality, and activities of eggshell gland antioxidant enzymes (catalase [CAT], glutathione peroxide [GSH-Px]), and ATPase (Na+/K+-ATPase, Ca2+-ATPase, and Mg2+-ATPase) deteriorated, and the activity of total nitric oxide synthase (T-NOS) and the level of malondialdehyde (MDA) increased significantly (P < 0.05). The histopathology and real-time quantitative PCR results showed that Cd induced endometrial epithelial cell proliferation accompanied by upregulation of the mRNA levels of progesterone receptor (PgR) and epidermal growth factor receptor (EGFR), downregulation of the mRNA levels of estrogen receptor α (ERα) and interleukin 6 (IL6), and inflammation of the eggshell gland accompanied by significantly increased expression of complement C3 and pro-inflammatory cytokine tumor necrosis factor α (TNFα) (P < 0.05). In addition, the ultrastructure of the eggshell showed that dietary supplementation with 7.5 mg/kg Cd increased the palisade layer and total thickness of the shell, but with the increase in dietary Cd supplementation (30 and 60 mg/kg) the thickness of the palisade layer and mammillary layer decreased significantly (P < 0.05), and the outer surface of the eggshell became rougher. Correspondingly, the expression of calbindin 1 (CALB1), ovocalyxin-32 (OCX-32), ovocalyxin-36 (OCX-36), osteopontin (SPP1), and ovocledidin-17 (OC-17) decreased significantly (P < 0.05) with increasing dietary Cd supplementation. Conclusively, the present study demonstrates that dietary supplementation with Cd negatively affects laying performance, egg quality, and eggshell deposition by disturbing the metabolism of eggshell glands in laying hens but has a positive effect on egg quality at low doses.

Dietary Cadmium Chloride Supplementation Impairs Renal Function and Bone Metabolism of Laying Hens.

This study was conducted to evaluate the toxic effects of cadmium (Cd) on the kidney function and bone development in laying hens. A total of 480 Hy-line laying hens aged 38 weeks were randomly allocated into five treatments, each of which included six replicates of 16 birds. The concentrations of Cd in the diets of the five groups were 0.47, 7.58, 15.56, 30.55, and 60.67 mg/kg. Results showed that serum calcium (Ca) levels decreased significantly in the 60.67 mg Cd/kg diet group (p < 0.05). The activities of serum alkaline phosphatase (ALP) and bone ALP (BALP) decreased significantly in the 15.56, 30.55 and 60.67 mg Cd/kg diet groups (p < 0.05). The levels of parathyroid hormone (PTH) increased significantly in the 30.55 and 60.67 mg Cd/kg diet groups, and the estradiol (E2), 1,25-(OH)2-D3 and calcitonin (CT) decreased significantly with the increase of dietary Cd supplementation (p < 0.05). Histological results presented enlargements of renal tubules and tubular fibrosis in the kidney and decreased trabecular bone in the tibia. Tartrate-resistant acidic phosphatase (TRAP) staining results of tibia showed that osteoclast was significantly increased at the relatively high dose of dietary Cd (p < 0.05). In addition, the renal function indicators of blood urea nitrogen (BUN), urea acid (UA), and creatinine were significantly increased in Cd supplemented groups compared with the control group (p < 0.05). Low dose Cd exposure induced antioxidant defenses accompanying the increase in activities of catalase (CAT), glutathione peroxidase (GSH-Px), and the levels of glutathione (GSH) in renal tissue. At the same time, with the increased Cd levels, the activities of CAT, GSH-Px decreased significantly, and the level of malondialdehyde (MDA) increased significantly (p < 0.05). The activities of Na+/K+-ATPase and Ca2+/Mg2+-ATPase decreased significantly in the relatively high levels of dietary Cd (p < 0.05). These results suggest that Cd can damage renal function and induce disorders in bone metabolism of laying hens.

Dietary addition of zinc-methionine influenced eggshell quality by affecting calcium deposition in eggshell formation of laying hens.

The present study explored the mechanism of Zn-methionine (Zn-Met) influencing eggshell quality of laying hens and investigated whether the mechanism was related to Ca deposition. Hyline grey layers (n 384, 38 weeks old) were divided into four groups: 0 mg Zn/kg, 40, 80 mg Zn/kg as Zn-Met, and 80 mg Zn/kg as zinc sulphate (ZnSO4). Eggshell quality, Zn contents, Zn-containing enzyme activities and expressions of shell matrix proteins in eggshell gland (ESG) were analysed. Zn-Met treatment at 80 mg/kg increased (P < 0·05) egg weight and eggshell strength throughout the experiments. The 80 mg/kg Zn-Met group (P < 0·05) had decreased mammillary knob width and larger relative atomic weight percentage of Ca, stronger signal intensity of Ca in linear distribution and the Ca was more evenly distributed in the transversal surface of eggshell. Zn contents (P < 0·001) in yolk and serum, Ca, albumin (Alb) levels in ESG as well as carbonic anhydrase (CA) activity in serum (P < 0·05) and mRNA levels of CA and Ca-binding protein-d28k (CaBP-D28k) (P < 0·001) in the 80 mg/kg Zn-Met group were the highest among all treatments. In conclusion, shell strength as one of eggshell qualities was mostly related to mammillary cone width in ultrastructure caused by the pattern of Ca deposition in eggshell formation. Also, the increase in Zn-Met-induced Ca deposition may be due to the increased Zn contents in serum and tissues, which were attributable to the increased CA concentrations in serum, Ca, Alb levels and up-regulated CA and CaBP-D28k mRNA levels in ESG.

Mercuric Chloride Induced Ovarian Oxidative Stress by Suppressing Nrf2-Keap1 Signal Pathway and its Downstream Genes in Laying Hens.

The present study evaluated the effects of mercury chloride (HgCl2) on follicular atresia rate, sex hormone secretion, and ovarian oxidative stress in laying hens. Antioxidant enzyme genes and the nuclear factor erythroid 2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) signal pathway were further studied to uncover the molecular mechanism. A total of 768 40-week-old Hy-Line Brown laying hens were randomly allocated to four treatments with eight pens per treatment and 24 hens of each pen. The birds were fed with four experimental diets containing graded levels of mercury (Hg) at 0.280, 3.325, 9.415, and 27.240 mg/kg, respectively. Results revealed that a positive relationship occurred between the accumulation of Hg in ovary and follicular atresia rate. Progesterone (P4) level significantly decreased in all Hg-treatment groups (P < 0.05), and follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were the lowest in the 27.240-mg/kg Hg group. Besides, the activities of catalase (CAT), superoxidative dismutase (SOD), glutathione reductase (GR), and glutathione (GSH) content were significantly decreased in all Hg-treatment groups (P < 0.05). Glutathione peroxidase (GSH-Px) activity significantly decreased, while malondialdehyde (MDA) content sharply increased in the 27.240-mg/kg Hg group (P < 0.05). In addition, there were positive relationships between antioxidant enzyme activities and antioxidant gene expressions or between antioxidant gene expressions and Nrf2 mRNA expression, while negative correlations occurred between Nrf2 and Keap1 at transcription and protein levels. It could be concluded that Hg induced ovarian function disorders and ovarian oxidative stress by means of impairing the Nrf2-Keap1 signal pathway in laying hens.

Curcumin Prevents Aflatoxin B1 Hepatoxicity by Inhibition of Cytochrome P450 Isozymes in Chick Liver.

This study was designed to establish if Curcumin (CM) alleviates Aflatoxin B1 (AFB1)-induced hepatotoxic effects and to determine whether alteration of the expression of cytochrome P450 (CYP450) isozymes is involved in the regulation of these effects in chick liver. One-day-old male broilers (n = 120) were divided into four groups and used in a two by two factorial trial in which the main factors included supplementing AFB1 (< 5 vs. 100 µg/kg) and CM (0 vs. 150 mg/kg) in a corn/soybean-based diet. Administration of AFB1 induced liver injury, significantly decreasing albumin and total protein concentrations and increasing alanine aminotransferase and aspartate aminotransferase activities in serum, and induced hepatic histological lesions at week 2. AFB1 also significantly decreased hepatic glutathione peroxidase, catalase, and glutathione levels, while increasing malondialdehyde, 8-hydroxydeoxyguanosine, and exo-AFB1-8,9-epoxide (AFBO)-DNA concentrations. In addition, the mRNA and/or activity of enzymes responsible for the bioactivation of AFB1 into AFBO-including CYP1A1, CYP1A2, CYP2A6, and CYP3A4-were significantly induced in liver microsomes after 2-week exposure to AFB1. These alterations induced by AFB1 were prevented by CM supplementation. Conclusively, dietary CM protected chicks from AFB1-induced liver injury, potentially through the synergistic actions of increased antioxidant capacities and inhibition of the pivotal CYP450 isozyme-mediated activation of AFB1 to toxic AFBO.

Prevention of Aflatoxin B1 Hepatoxicity by Dietary Selenium Is Associated with Inhibition of Cytochrome P450 Isozymes and Up-Regulation of 6 Selenoprotein Genes in Chick Liver.

BACKGROUND: The involvement of cytochrome P450 (CYP450) isozymes and the selenogenome in selenium-mediated protection against aflatoxin B1 (AFB1)-induced adverse effects in broilers remains unclear. OBJECTIVE: This study was designed first to determine whether selenium could reduce AFB1-induced hepatotoxic effects and then to determine whether these effects were due to changes in the CYP450 isozymes and selenogenome expression in the liver of chicks. METHODS: Male avian broilers (aged 120 d) were allocated to 4 groups with 5 replicates of 6 birds to be included in a 2-by-2 factorial trial in which the main factors included supplementation of AFB1 (<5 compared with 100 µg/kg) and selenium (0.2 compared with 0.5 mg/kg) in a corn/soybean-based diet for 4 wk. Serum biochemistry, hepatic histology, and mRNA and/or activities of hepatic antioxidant enzymes, CYP450 isozymes, and 26 selenoproteins were analyzed at week 2 and/or 4. RESULTS: Administration of AFB1 induced liver injury, decreasing (P < 0.05) total protein and albumin concentrations by 33.3-43.8% and increasing (P < 0.05) alanine aminotransferase and aspartate aminotransferase activities by 26.0-33.8% in serum, and induced hepatic necrosis and bile duct hyperplasia at week 2. AFB1 also decreased (P < 0.05) hepatic activities of glutathione peroxidase (GPX), thioredoxin reductase (TXNRD), and catalase, and the glutathione concentration by 13.1-59.9% and increased (P < 0.05) malondialdehyde, 8-hydroxydeoxyguanosine and exo-AFB1-8,9-epoxide (AFBO) DNA concentrations by 17.9-1200%. In addition, the mRNA and activity of enzymes responsible for the bioactivation of AFB1 into AFBO, which included CYP450 A1, 1A2, 2A6, and 3A4, were significantly induced (P < 0.05) by 29.2-271% in liver microsomes after 2-wk exposure to AFB1. These alterations induced by AFB1 were prevented by selenium supplementation. Dietary selenium supplementation increased (P < 0.05) mRNA and/or activities of 6 selenoprotein genes (Gpx3, Txnrd1, Txnrd2, Txnrd3, iodothyronine deiodinase 2, and selenoprotein N) in the liver of AFB1-treated groups at week 2. CONCLUSIONS: Dietary selenium protected chicks from AFB1-induced liver injury, potentially through the synergistic actions of inhibition of the pivotal CYP450 isozyme-mediated activation of AFB1 to toxic AFBO, and increased antioxidant capacities by upregulation of selenoprotein genes coding for antioxidant proteins.